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ABSTRACT: Background and Objective: B7-H1, a member of B7 family, is expressed in tumor cells and has emerged as an important immune modulator capable of suppressing host immunity by inhibiting T cells function. This study was to probe into the correlation between the expression level of B7-H1 protein in pancreatic carcinoma tissues and clinicopathological characteristics and prognosis. Methods: The expression of B7-H1 was measured in 40 cases of pancreatic carcinoma tissues and 10 cases of normal corresponding paracarcinoma tissues by immunohistochemistry. The relationship between the expression level of B7-H1 and clinicopathological characteristics and survival was analyzed. Results: The positive rate of B7-H1 was significantly higher in the tumor tissues [45.00% (18/40)] than in the normal corresponding paracarcinoma tissues [0(0/10)] (P<0.05); moreover, B7-H1 expression was significantly associated with the staging of tumor and preoperative serum CA19-9 level (P<0.05). The multivariate cox proportional hazards regression analysis of prognostic factors for overall survival and relapse-free survival showed that the expression of B7-H1 was an independent factor for poor prognosis. Conclusion: B7-H1 protein was expressed in human pancreatic carcinoma tissues, and was associated with the prognosis.
Ai zheng = Aizheng = Chinese journal of cancer 12/2009; 28(12):1328-32.
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ABSTRACT: Cancer patients have a poor immune response and chemotherapy could deteriorate their immune system further. Reasonable immune therapy is an important adjuvant remedy for tumors. This study was to monitor the changes of T-cell phenotypes in peripheral blood and interleukin-2 (IL-2) concentration in plasma in digestive tract cancer patients before and after chemotherapy.
The proportions of CD3+, CD4+, CD8+, CD4+CD28+, CD8+CD28+ and CD4+CD25+ T cells in peripheral blood of 104 patients with advanced digestive tract cancer, hospitalized from Sep. 2005 to Apr. 2006, were detected by flow cytometry (FCM). The concentration of IL-2 in plasma was measured by ELISA.
The proportions of CD4+, CD4+CD28+, CD8+CD28+, CD4+CD25+ T cells and ratio of CD4/CD8 were (36.52+/-3.85)%, (32.87+/-4.98)%, (6.87+/-3.11)%, (9.68+/-3.42)% and 0.98+/-0.17 in digestive tract cancer patients, and (45.23+/-9.20)%, (40.12+/-5.85)%, (15.8+/-4.50)%, (5.67+/-2.90)% and 1.43+/-0.12 in healthy subjects. In the patients with response to chemotherapy, the proportions of CD4+CD28+ and CD8+CD28+ T cells and ratio of CD4/CD8 were (22.93+/-3.98)%, (7.08+/-1.23)% and 0.90+/-0.22 before chemotherapy, and (28.25+/-4.03)%, (12.10+/-3.45)% and 1.24+/-0.22 at 3 weeks after chemotherapy. In the patients with no response to chemotherapy, the proportions of CD4+CD28+, CD8+CD28+ and CD4+CD25+ T cells were (24.08+/-4.02)%, (6.35+/-1.23)% and (8.20+/-2.34)% before chemotherapy, and (16.45+/-3.27)%, (3.20+/-0.82)% and (20.34+/-3.69)% at 3 weeks after chemotherapy.
The immunosuppression of digestive tract cancer patients would be enhanced early (about 1-2 weeks) after intravenous chemotherapy. The immunity of the patients with response to chemotherapy would be improved at 3 weeks after chemotherapy; while the immunity of the patients with no response to chemotherapy would not change, or even be suppressed.
Ai zheng = Aizheng = Chinese journal of cancer 04/2008; 27(4):418-24.
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ABSTRACT: To prepare novel anti-OX40L functional monoclonal antibodies and characterize their distinct biological functions.
Routine immunization of BALB/c mice by a tansfected cell line L929/OX40L expressing high level of OX40L as antigens. Then fuse the immunized spleen with Sp2/0, a kind of myloma, and screen the positive clones by FCS with L929/OX40L as a positive control.After acquisition of the hybidomas secreting anti-OX40L mAb, investigation of their biological activities by Western blot, rapid isotyping analysis, karyotype analysis, competitive inhibition test, indirect immunofluorescence and MTT incorporation assay were followed.
Obtain two stable hybridomas, 4D6 and 5C2, which could continuously secret specific anti-OX40L monoclonal antibodies. The following biological activity studies showed that these monoclonal antibodies could both recognize the natural OX40L expressed on the mature DC, especially the OX40L on the several leukemia cell lines, such as Jurkat, SHI-1, U937 etc. Furthermore, they could also suppress the proliferation of SHI-1 in vitro.
Two hybridomas secreting anti-OX40L monoclonal antibodies continuously and steadily have been established. These monoclonal antibodies could specifically recognize human OX40L and effect the proliferation of leukemia cell line SHI-1 through OX40/OX40L costimulatory signals.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 09/2007; 23(8):757-60.
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ABSTRACT: To explore the biofunctions of human B7-H4 generated from prokaryotic system.
The gene of human B7-H4 extracellular region (IgV-like and IgC-like domains) was obtained by PCR from human cDNA FLJ22418 and then inserted into the prokaryotic expression vector pGEX-5X-3 expressing glutathione s-transferase (GST) fusion protein. After being identified by restriction enzyme digestion and sequencing, the recombinant vector was transferred into host strain E coli BL21-RIL(DE3). A 47 kDa fusion protein (GST/hB7-H4) was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified by standard methods reported in the prokaryotic system. The inhibitory effect of GST/hB7-H4 on proliferation of T cells was observed in vitro by CD3mAb activated T-cell culturing system and [(3)H]-thymidine incorporation assay. The concentrations of interleukin-2 and iterferon-g in the supernatants of T cells were determined by ELISA.
We successfully constructed the method for high-level expression and purification of the hB7-H4 extracellular domain as GST fusion protein from E coli. The GST/hB7-H4 fusion protein produced in bacteria had obvious biological activity to inhibit T-lymphocyte proliferation and IL-2 secretion.
The prokaryote expression system could be used to generate hB7-H4 protein with natural spatial conformations and biological functions, which provided an efficient and economical way for the preparation of this target protein.
Acta Pharmacologica Sinica 07/2006; 27(6):741-6. · 1.95 Impact Factor
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ABSTRACT: Aim: To explore the biofunctions of human B7-H4 generated from prokaryotic system. Methods: The gene of human B7-H4 extracellular region (IgV-like and IgC-like domains) was obtained by PCR from human cDNA FLJ22418 and then inserted into the prokaryotic expression vector pGEX-5X-3 expressing glutathione s-transferase (GST) fusion protein. After being identified by restriction enzyme digestion and sequencing, the recombinant vector was transferred into host strain E coli BL21-RIL(DE3). A 47 kDa fusion protein (GST/hB7-H4) was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified by standard methods reported in the prokaryotic system. The inhibitory effect of GST/hB7-H4 on proliferation of T cells was observed in vitro by CD3mAb activated T-cell culturing system and [3H]-thymidine incorporation assay. The concentrations of interleukin-2 and iterferon-g in the supernatants of T cells were determined by ELISA. Results: We successfully constructed the method for high-level expression and purification of the hB7-H4 extracellular domain as GST fusion protein from E coli. The GST/hB7-H4 fusion protein produced in bacteria had obvious biological activity to inhibit T-lymphocyte proliferation and IL-2 secretion. Conclusion: The prokaryote expression system could be used to generate hB7-H4 protein with natural spatial conformations and biological functions, which provided an efficient and economical way for the preparation of this target protein.
Acta Pharmacologica Sinica 06/2006; 27(6):741 - 746. · 1.95 Impact Factor
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ABSTRACT: Although the roles of CD40 in B cells have been intensively studied, little is known on the function of CD40 in lung cancer cell lines. This study was to investigate biological effects of soluble CD40 ligand (sCD44L) on lung cancer cell line A549 (CD40 positive), and its possible mechanism.
A549 cells were co-incubated with sCD40L, cell proliferation was detected by MTT assay and 3H-TdR incorporation method. Immunofluorescence technique and flow cytometry (FCM) were used to evaluate changes in cell phenotypes and cell cycle. Cell apoptosis, and expression changes of Bcl-2 and Bax were observed by FCM, reverse transcriptase-polymerase chain reaction (RT-PCR), and Western blot.
Compared with control cells, proliferation of A549 cells co-incubated with sCD40L was inhibited (P< 0.05). Positive rates of cell surface molecules, CD49e, CD54, TNFRI, and CD95L, in A549 cells co-incubated with sCD40L for 72 h were (61.2+/-4.8)%, (31.2+/-6.1)%,(42.7+/-5.9)%, and (38.2+/-3.4)%, respectively, while those in control cells were (34.7+/-2.1)%, (7.1+/-1.6)%, (15.2+/-4.1)%, and (10.1+/-2.3)%, respectively (P< 0.05). However, positive rate of TNFRII in A549 cells co-incubated with sCD40L[(8.7+/-0.8)%] was lower than that in control cells [(58.1+/-3.6)%] (P< 0.05). G1 phase of A549 cells treated with sCD40L for 72 h was (76.0+/-9.1)%, more than that of control cells [(56.7+/-6.9)%], while S phase of sCD40L-treated A549 cells [(10.3+/-5.7)%] was less than that of control cells [(32.7+/-5.5)%]. No significant apoptosis of A549 cells was observed after co-incubated with sCD40L for 72 h, but Bax expression was up-regulated.
sCD40L may inhibit cell proliferation, cause changes in phenotype and cell cycle of A549 cells, and alter expression of apoptosis-associated gene, such as Bax.
Ai zheng = Aizheng = Chinese journal of cancer 11/2004; 23(11):1278-82.
